Ocular allergy treatments

ABSTRACT

Compositions, kits and methods for the treatment or prevention of ocular allergies and inflammation and the symptoms thereof comprising alcaftadine or a pharmaceutically acceptable salt thereof.

RELATED APPLICATION

This application is a continuation-in-part of a non-provisional filingU.S. application patent Ser. No. 11/688,016, which claims priority froma provisional filing, U.S. App. Pat. Ser. No. 60/788185, entitled“Ocular Allergy Treatments,” which was filed on, Mar. 31, 2006.

FIELD OF THE INVENTION

This invention is directed to the treatment or prevention of ocularconditions. More specifically, the invention is directed to compositionsof alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid, as well as methods for using such compositions for the treatmentor prevention of ocular inflammation and allergies.

BACKGROUND

Allergic disorders of the ocular surface include a wide variety ofpathological conditions including Seasonal Allergic Conjunctivitis(“SAC”), Perennial Allergic Conjunctivitis (“PAC”), VernalKeratoconjunctivitis and Atopic Keratoconjunctivitis. It is estimatedthat over 20% of the general population suffer from some form of ocularallergy. Of those, approximately 90% suffer from either SAC, PAC orboth.

The ocular allergic reaction is an IgE-dependent (Type I)hypersensitivity inflammatory response that most commonly affects adultsbetween 20 and 40 years of age. In susceptible individuals, initialexposure of allergen to the ocular surface stimulates the production ofallergen specific immunologic antibodies (IgE). IgE then binds to themembrane bound FcεR-1 receptor of naive mast cells in the ocular mucosa.The mast cell is a granulocyte, containing a number of preformedmediators, including histamine and proteoglycans. Once the mast cell isactivated, newly formed chemical mediators are formed, which includeprostaglandin D2, leukotrienes, and platelet aggregating factor.Subsequent exposure of allergen to the IgE coated mast cells leads tothe release of preformed, as well as newly formed, mediators containedwithin the granules of the mast cell.

The clinical symptoms of allergic conjunctivitis include itching,redness, swelling of the eyelid, chemosis and tearing. Histamine is theprimary mediator in the allergic response. After mast celldegranulation, histamine binds to receptors located in the conjunctiva.The binding of histamine to H1 receptors on nerve cells induces itching.Activation of H1 and H2 receptors on the vaso-endothelium inducesvasodilatation and increases vascular permeability facilitating themigration of inflammatory mediators, such as IL-1α and IL-1β, into theblood vessel and the subsequent recruitment of leukocytes into theconjunctival tissue. Activation of the histamine receptors leads toocular hyperemia, chemosis, lid swelling and exudation of fluid fromblood vessels into the surrounding tissue, which in turn causesinflammation. The chemotaxis of leukocytes such as eosinophils andneutrophils into the conjunctival tissue in turn leads to further tissuedamage.

Historically, antihistamines have been the mainstay for treatment ofocular allergic disease. These therapies vary in potency, specificityand duration of action. First generation anti-histamines such aspheniramine and antazoline are known for their rapid onset of action.Unfortunately, these compounds also cause ocular discomfort and theirefficacy diminishes after only a few hours. Second-generation H1antagonists such as levocabastine and emadastine present less oculardiscomfort and have a somewhat longer duration of action. However, thesecompounds have limited anti-inflammatory effects, and do little toinhibit the late-phase components of the inflammatory response.

Currently, the most effective therapies for the management of ocularallergy are drugs such as olopatadine, ketotifen and azelastine, whichhave both anti-histaminic and mast cell stabilizing properties. Thesetherapies are generally well tolerated and their effects can last up to8 to 12 hours. Although reported to be superior to compounds that effectonly a single component of the allergic response, these compounds oftenfail to provide relief more than one ocular allergy symptoms.

A drug's affect on ocular redness, chemosis and eyelid swelling offers asignificant improvement over existing therapies. Additionally, since themajority of newer ophthalmic anti-allergic agents have limited durationsof action, twice daily dosing is required. A topical preparation with alonger duration of action will be advantageous because it may beinstilled once daily. Thus, new therapies that can offer advantages inareas such as efficacy and duration of action, while offering similarsafety profiles, are needed. The instant invention is directed to theseand other objectives.

DETAILED DESCRIPTION OF THE INVENTION

The invention includes methods of treating or preventing ocular allergyby administering alcaftadine to the eye of a patient. The inventionsdescribed herein are based at least in part on the surprising discoverythat alcaftadine treats or prevents a number of different symptoms ofocular allergy that make it especially useful for the treatment orprevention of ocular allergy. The methods, ophthalmic compositions, andkits of the present invention alleviate clinical symptoms of ocularallergy and ocular inflammation with minimal systemic absorption of theactive drug. This unusual combination of properties, together with anexcellent safety profile and tolerability when formulated for topicaladministration to the eye, makes the drug especially useful for thetreatment or prevention of ocular allergy. Specifically, the inventionincludes a method of treating or preventing a clinical symptom of ocularallergy, comprising administering to the eye of a patient an effectiveamount of alcaftadine, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof.

Alcaftadine, also known by the chemical name6,11-dihydro-11-(1-methyl-4-piperidinylidene)-5H-imidazo [2,1-b][3]benzazepine-3-carboxaldehyde, has the following chemical formula:

The compound and methods for its preparation are disclosed in U.S. Pat.No. 5,468,743, which is incorporated herein by reference in its entiretyfor all purposes. The preferred methods and ophthalmic compositions ofthe invention contain the alcaftadine compound of Formula I, but mayalternatively be present as an alcaftadine salt. Pharmaceuticallyacceptable salts of alcaftadine can be formed from organic and inorganicacids. Suitable acids include, but are not limited to, acetic,4-acetamido benzoic acid, benzenesulfonic, camphorsulfonic, citric,2,3:4,6-di-O-isopropylidene-2-keto-L-gulonic acid monohydrate, formic,fumaric, hydrochloric, hydrobromic, lactic, maleic, L-(−)malic, malic,malonic, mandelic, methanesulfonic, naphthalenesulfonic, nitric, oxalic,phthalic, phosphoric, propionic, DL-pyroglutamic, saccharin, salicyclic,succinic, sulfuric, tartaric, trifluoro acetic, L-(+)tartaric, andtoluenesulfonic acids.

As used herein the terms “ocular allergy” refers to an allergic disorderof the ocular surface caused by pathogenic allergens. Allergicconjunctivitis is the preferred ocular allergy and includes a widevariety of pathological conditions including Seasonal AllergicConjunctivitis (“SAC”), Perennial Allergic Conjunctivitis (“PAC”),Vernal Keratoconjunctivitis and Atopic Keratoconjunctivitis.

“Clinical symptoms” of ocular allergy include but are not limited toocular itching, ocular redness, swelling of the eyelids, chemosis,tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasalpruritis and ear/palate pruritis, and sneezing. It is preferred that themethods of the invention treat or prevent at least two clinicalsymptoms, more preferably at least three, even more preferably more thatfour. For example, the methods of the invention treat or prevent atleast one of the following clinical symptoms associated with allergicconjunctivitis ocular itching, ocular redness, chemosis, tearing,swelling of lid nasal congestion, or rhinorrhea. Preferably the methodsof the invention treat or prevent, ocular itching and ocular redness;treat or prevent ocular itching, ocular redness, and chemosis; treat orprevent ocular itching, ocular redness, chemosis, and tearing; treat orprevent ocular itching, ocular redness, chemosis, tearing, and swellingof the lid; treat or prevent ocular itching, ocular redness, chemosis,tearing, swelling of the lid, and nasal congestion; treat or preventocular itching, ocular redness, chemosis, tearing, swelling of the lid,nasal congestion, and rhinorrhea; treat or prevent nasal congestion andrhinorrhea.

The term “patient,” as used herein, refers to animals, includingmammals, preferably humans. The “effective amount” of alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof, is the amount the substancerequired to treat or prevent the symptoms of ocular allergy. Theeffective amount may vary from patient to patient depending upon theability of alcaftadine, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof, alone or in combination with one or more combination drugs toelicit a desired response in the patient. Other factors determining theeffective amount will include, but are not limited to the disease stateor severity of the condition to be alleviated, hormone levels, age, sex,weight of the patient, the state of being of the patient, and theseverity of the pathological condition being treated, concurrentmedication or special diets then being followed by the particularpatient, and other factors which those ordinarily skilled in the artwill recognize, with the appropriate dosage ultimately being at thediscretion of the attending physician. Dosage regimens may be adjustedto provide the improved therapeutic response. An effective amount isalso one in which any toxic or detrimental effects of the components areoutweighed by the therapeutically beneficial effects. It is preferredthat for most patients a 50 μL drop of a 0.25% ocular solution contains0.125 mg of alcaftadine. Assuming that 100% of drug is systemicallyabsorbed, a 70 kg person, using the eye drops bilaterally, meaning ineach eye, once daily, would be exposed to a dose of 0.25 mg/d, or 3.57μg/kg per day. It is reasonable to assume that the actual systemicexposure will be lower, since it is likely that not all of the amountwill be absorbed. It is preferred that the effective amount ofalcaftadine, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof, isbetween less than about 0.25 mg and greater than or equal to about 0.015mg, more preferably, between about 0.030 mg and about 0.14 mg, morepreferably between about 0.075 mg and about 0.125 mg.

The term “pharmaceutically acceptable” as used herein refers tomaterials that are generally not toxic or injurious to a patient whenused with alcaftadine, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof, the present invention, including when the alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof, , is formulated asophthalmic compositions, as defined herein.

Alcaftadine, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof, may beadministered to the patient by any route of administration capable ofdelivering the drug to the eye of the patient, in any pharmaceuticallyacceptable dosage form. Thus, the drug may be administered to thepatient in the form of an ophthalmic composition, as defined herein, orany other formulation, device or mechanism suitable for the short termor long term delivery of an effective amount of the drug to thepatient's eye. The drug may be administered to the patient in anophthalmic inserts containing or coated with alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof, including but not limited tocontact lenses, punctal plugs, or, ocular inserts. In preferred methodsof the present invention, the drug is administered topically in the formof an ophthalmic composition selected from the group consisting ofophthalmic solutions or suspensions (i.e., eye drops), ophthalmicointments, or ophthalmic gels.

Further, the invention includes a method of treating or preventing aclinical symptom of ocular inflammation, comprising administering to theeye of a patient an effective amount of alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms alcaftadine,clinical symptom, patient, pharmaceutically acceptable, pharmaceuticallyacceptable salts, and effective amount all have their aforementionedmeanings and preferred ranges. The term ocular inflammation refers toinflammation of any part of the anterior portion of the eye. Such ocularinflammation may be caused by any of the following or any combination ofthe following dry eye, contact lens wear, bacterial infections, fungalinfections, or viral infections. The preferred causes of ocularinflammation are bacterial infections or viral infections.

In addition the invention includes a method of treating or preventing amechanistic symptom associated with ocular allergy or ocularinflammation comprising administering to the eye of a patient aneffective amount of alcaftadine, its pharmaceutically acceptable salts,its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof. The terms alcaftadine, patient pharmaceutically acceptable,pharmaceutically acceptable salts, effective amount, ocular allergy, andocular inflammation all have their aforementioned meanings and preferredranges. “Mechanistic symptoms” are cellular reactions that either elicitor suppress symptoms of a disease state such as ocular allergy or ocularinflammation. Mechanistic symptoms include but are not limited tovascular leakage, a reduction in the integrity of the conjunctivalepithelial tight junctions, modulation of the H₄ receptor, and mast celldegradation. The preferred methods of the invention treat or prevent atleast two mechanistic symptoms, more preferably treat or prevent atleast three mechanistic symptoms, even more preferably treat or preventat least four mechanistic symptoms. For example the preferred methodstreat or prevent vascular leakage, and a reduction in the integrity ofthe conjunctival epithelial tight; treat or prevent vascular leakage, areduction in the integrity of the conjunctival epithelial tightjunctions, and modulation of theH₄ receptor; treat or prevent vascularleakage, a reduction in the integrity of the conjunctival epithelialtight junctions, modulation of theH₄ receptor, and mast celldegradation.

Further still, the invention includes a method of treating or preventinga nasal symptom of ocular allergy, comprising administering to the noseof a patient an effective amount of alcaftadine, its pharmaceuticallyacceptable salts, its N-oxides, hydrates, solvates, polymorphs,pro-drugs, or mixtures thereof. The terms alcaftadine, patient,pharmaceutically acceptable, pharmaceutically acceptable salts, ocularallergy, and effective amount all have their aforementioned meanings andpreferred ranges. “Nasal symptoms” of allergy are a subset of clinicalsymptoms as defined above and include nasal inflammation, nasalcongestion, rhinorrhea, nasal pruritis, and sneezing. The preferrednasal symptoms are rhinorrhea and nasal congestion.

Alfcaftadine may be administered to the patient in the form of anophthalmic composition, as defined herein, or any other formulation,device or mechanism suitable for the short term or long term delivery ofan effective amount of the drug to the patient's nose, preferably thepatient's nostrils. In the preferred methods of the present invention,alcaftadine, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof, isadministered to the patient's nostrils topically in the form of anophthalmic composition selected from the group consisting of ophthalmicsolutions or suspensions (i.e., nasal drops and spray), ophthalmicointments, or ophthalmic gels (as defined herein).

Further the invention includes a method of treating or preventing aclinical symptom of ocular allergy, comprising administering to the eyeof a patient an ophthalmic composition comprising alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms alcaftadine,clinical symptom, ocular allergy, patient, pharmaceutically acceptable,and pharmaceutically acceptable salts, all have their aforementionedmeanings and preferred ranges.

As used herein the term “ophthalmic composition” refers to anypharmaceutically acceptable formulation, delivery device, mechanism orsystem suitable for administration to the eye. The term “ophthalmiccompositions” includes but are not limited to solutions, suspensions,gels, ointments, contact lenses, implants, sprays, depots or any othertype of formulation, device or mechanism suitable for short term or longterm delivery of alcaftadine, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof, to the eye. In contrast to oral or injectable formulations,ophthalmic compositions exhibit specific technical characteristicsassociated with their application to the eyes, including the use ofpharmaceutically acceptable ophthalmic vehicles that avoid inducingvarious reactions such as, for example, irritation of the conjunctivaand cornea, closure of the eyelids, secretion of tears and painfulreactions. Preferred ophthalmic compositions according to the inventionare advantageously in the form of ophthalmic solutions or suspensions(i.e., eye drops), ophthalmic ointments, or ophthalmic gels containingalcaftadine, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.Depending upon the particular form selected, the compositions maycontain various additives such as buffering agents, isotonizing agents,solubilizers, preservatives, viscosity-increasing agents, chelatingagents, antioxidizing agents, and pH regulators.

Examples of suitable preservatives include, but are not limited tochlorobutanol, sodium dehydroacetate, benzalkonium chloride, cetylpyridinium chloride, phenethyl alcohol, parahydroxybenzoic acid esters,and benzethonium chloride. The viscosity-increasing agents may beselected, for example, from methylcellulose, hydroxyethylcellulose,carboxymethylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol,carboxymethylcellulose, chondroitin sulfate, and salts thereof. Suitablesolubilizers include, but are not limited to, polyoxyethylenehydrogenated castor oil, polyethylene glycol, polysorbate 80, andpolyoxyethylene monostearate. Typical chelating agents include, but arenot limited to, sodium edetate citric acid, stabilizing agents asdefined in U.S. App. Pat. No. 60/783,557 filed on, Mar. 17, 2006,entitled “Methods for Stabilizing Oxidatively Unstable PharmaceuticalCompositions” and its corresponding non-provisional filing which arehereby incorporated by reference in their entirety. The stabilizersinclude, but are not limited to for example, sodium edetate and sodiumhydrogen sulfite.

Useful pH regulators are commonly selected, for example, from sodiumhydroxide, potassium hydroxide, sodium carbonate, citric acid,phosphoric acid, acetic acid, and hydrochloric acid. The pH of theophthalmic compositions may range from about 5 to about 8, morepreferably from about 6.5 to about 7.5. Even more preferably, the pH ofthe ophthalmic compositions is about 7.0. Useful buffers include, butare not limited to borate buffers, phosphate buffers, carbonate buffers,and acetate buffers. The concentration of buffer in the ophthalmiccompositions may vary from about 1 mM to about 150 mM or more, dependingon the particular buffer chosen. Preferably, the concentration of bufferis less than 100, more preferably from about 1 mM to about 25 mM, with aconcentration of about 1 mM to about 20 mM more preferred.

As used herein, the term “vehicle” is intended to include any carrier,diluent or excipient suitable for ophthalmic use. “Excipient” refers toan ingredient that provides one or more of bulk, imparts satisfactoryprocessing characteristics, helps control the dissolution rate, andotherwise gives additional desirable characteristics to thecompositions. Included within this term, inter alia, are compounds wellknown to those of ordinary skill in the art, as described, for example,in the Handbook of Pharmaceutical Excipients, (American PharmaceuticalAssociation, Washington, D.C. and Pharmaceutical Press, London, England,4^(th) ed. 2003), incorporated herein by reference in its entirety. Inparticular, the excipients are selected such that the ophthalmiccomposition does not trigger a secretion of tears that will entrain theactive ingredient. Acceptable excipients are well known to a personskilled in the art, who will know how to select them depending on thedesired formulation.

When concentrations, amounts, percentages, and other numerical data areexpressed or presented herein in a range format, it is to be understoodthat such a range format is used merely for convenience and brevity andthus are to be interpreted flexibly to include not only the numericalvalues explicitly recited as the limits of the range, but also toinclude each of the individual numerical values or sub-rangesencompassed within that range as if each numerical value and sub-rangeis explicitly recited. As an illustration, a concentration range of“about 1 weight % to about 10 weight %” is to be interpreted to includenot only the explicitly recited concentration of about 1 weight % toabout 10 weight %, but also individual concentrations and the sub-rangeswithin the indicated range. Thus, included in this numerical range areindividual concentrations such as 2 weight %, 5 weight %, and 8 weight%, and sub-ranges such as from 1 weight % to 3 weight %, from 5 weight %to 9 weight % and so forth. As used herein, the term “about” means plusor minus approximately ten percent of the indicated value, such that“about 50% by weight” indicates approximately 45% to 55% by weight.

Typically, the concentration of alcaftadine in the ophthalmiccompositions of the present invention will be from about 0.005% byweight to about 10.0% by weight, with concentrations of from about 0.005to about 0.4% being preferred, and concentrations of about 0.1% to about0.35% being particularly preferred. A 50 μL drop of a 0.25% ocularsolution contains 0.125 mg of alcaftadine. Assuming that 100% of drug issystemically absorbed, a 70 kg person, using the eye drops bilaterally,meaning in each eye, once daily, would be exposed to a dose of 0.25mg/d, or 3.57 μg/kg per day. It is reasonable to assume that the actualsystemic exposure will be lower, since it is likely that not all of theamount will be absorbed.

Further, still the invention includes a method of treating or preventinga mechanistic symptom of ocular allergy, comprising administering to theeye of a patient an ophthalmic composition comprising alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms alcaftadine,mechanistic symptom, patient, pharmaceutically acceptable, andpharmaceutically acceptable salts, all have their aforementionedmeanings and preferred ranges.

Still further, the invention includes a method of treating or preventinga clinical symptom of ocular inflammation, comprising administering tothe eye of a patient an ophthalmic composition comprising alcaftadine,its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms alcaftadine,clinical symptom, patient, pharmaceutically acceptable, andpharmaceutically acceptable salts, ocular inflammation, and ophthalmiccomposition all have their aforementioned meanings and preferred ranges.

Yet still further, the invention includes a method of treating orpreventing a mechanistic symptom of ocular inflammation, comprisingadministering to the eye of a patient an ophthalmic compositioncomprising alcaftadine, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof. The terms alcaftadine, mechanistic symptom, patient,pharmaceutically acceptable, and pharmaceutically acceptable salts,ocular inflammation, ophthalmic composition all have theiraforementioned meanings and preferred ranges.

Further, the invention includes an ophthalmic composition comprisingalcaftadine, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof. Theterms ophthalmic composition, alcaftadine, pharmaceutically acceptablesalts all have their aforementioned meanings and preferred ranges. It ispreferred that said ophthalmic composition further comprise a vehicle asdefined herein.

Yet further still, the invention includes use of alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof in the preparation of amedicament for the treatment or prevention of a clinical symptom ofocular allergy. The terms alcaftadine, clinical symptom, ocular allergypharmaceutically acceptable salts all have their aforementioned meaningsand preferred ranges

Yet still further, the invention includes use of alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof in the preparation of amedicament for the treatment or prevention of a clinical symptom ofocular inflammation. The terms alcaftadine, clinical symptom, ocularinflammation pharmaceutically acceptable salts all have theiraforementioned meanings and preferred ranges.

Yet further still, the invention includes use of alcaftadine, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof in the preparation of amedicament for the treatment or prevention of a mechanistic symptom ofocular allergy or ocular inflammation. The terms alcaftadine,mechanistic symptom, ocular allergy, ocular inflammation,pharmaceutically acceptable salts all have their aforementioned meaningsand preferred ranges

Still further, the invention includes a kit comprising an ophthalmiccomposition comprising alcaftadine, its pharmaceutically acceptablesalts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, ormixtures thereof, contained within a container prepared from apharmaceutically acceptable packaging material. The terms ophthalmiccomposition, alcaftadine, pharmaceutically acceptable salts all havetheir aforementioned meanings and preferred ranges. Pharmaceuticallyacceptable packaging materials include but are not limited to lowdensity polyethylene (“LDPE”), high density polyethylene (“HDPE”),polypropylene, polystyrene, polycarbonate, polyesters (such aspolyethylene terephthalate and polyethylene naphthalate), nylon,poly(vinyl chloride), poly(vinylidine chloride),poly(tetrafluoroethylene) and other materials known to those of ordinaryskill in the art. Flexible bottles prepared from LDPE or HDPE areparticularly preferred. Commercial sources of such materials include butare not limited to DuPont 20 Series specialty polyethylene, manufacturedby DuPont, Tenite Polyethylene 1830F Natural, manufactured by EastmanChemical Company, Purell 1840 Polyethylene, manufactured by Basell. Theparticularly preferred material is DUPONT™ 20-6064 (E. I. du Pont deNemours and Company, Wilmington, Del.), a preferred LDPE packagingmaterial, is commonly used for preparing flexible dropper bottlescontaining ophthalmic compositions by an injection blow molding process,and is approved for such use by the U.S. Food and Drug Administration.The kits may contain multiple doses of ophthalmic compositionscontaining alcaftadine or single use doses of alcaftadine.

Prior to filling, such bottles are routinely sterilized by gammairradiation or with ethylene oxide gas, by methods widely known to thoseskilled in the art. Applicants have surprisingly found, however, that itis preferable to sterilize LDPE bottles with ethylene oxide gas, insteadof with gamma radiation, as bottles sterilized with gamma radiation mayexhibit decreased stability of the active ingredient.

6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (“CAS #147083-93-0”) has the following chemical formula

The compound of Formula II was disclosed in U.S. Pat. No. 5,468,743.Preferred methods and ophthalmic compositions may contain Formula II asdepicted, but Formula II may be present in the methods and ophthalmiccompositions as its pharmaceutically acceptable salts, N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.

Specifically the invention includes a method of treating or preventing aclinical symptom of ocular allergy, comprising administering to the eyeof a patient an effective amount of a compound of Formula II, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms a compound ofFormula II, clinical symptom, ocular allergy, patient, pharmaceuticallyacceptable, and pharmaceutically acceptable salts, all have theiraforementioned meanings and preferred ranges. The term “effectiveamount” of a compound of Formula II, its pharmaceutically acceptablesalts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, ormixtures thereof is the amount of this substance required to treat orprevent the symptoms of ocular allergy. As described earlier inreference to the effective amount of alcaftadine, its pharmaceuticallyacceptable salts, its N-oxides, hydrates, solvates, polymorphs,pro-drugs, or mixtures thereof, the effective amount of a compound ofFormula II, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof, may varyfrom patient to patient depending upon the ability of a compound ofFormula II, its pharmaceutically acceptable salts, N-oxides, hydrates,solvates, polymorphs, pro-drugs, and mixtures thereof, alone or incombination with one or more combination drugs to elicit a desiredresponse in a patient. Factors used to determine the effective amountare known to those of ordinary skill and some those factors arementioned herein. It is preferred that the effect amount of a compoundof Formula II is between less than about 0.25 mg and greater than orequal to about 0.015 mg, more preferably, between about 0.030 mg andabout 0.14 mg, more preferably between about 0.075 mg and about 0.125mg.

Further, the invention includes a method of treating or preventing aclinical symptom of ocular inflammation, comprising administering to theeye of a patient an effective amount of a compound of Formula II, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms a compound ofFormula II, clinical symptom, ocular inflammation patient,pharmaceutically acceptable, pharmaceutically acceptable salts, andeffective amount all have their aforementioned meanings and preferredranges.

In addition the invention includes a method of treating or preventing amechanistic symptom associated with ocular allergy or ocularinflammation comprising administering to the eye of a patient aneffective amount of a compound of Formula II, its pharmaceuticallyacceptable salts, N-oxides, hydrates, solvates, polymorphs, pro-drugs,or mixtures thereof. The terms a compound of Formula II, patientpharmaceutically acceptable, pharmaceutically acceptable salts,effective amount, mechanistic symptoms, ocular allergy, and ocularinflammation all have their aforementioned meanings and preferredranges.

Further still, the invention includes a method of treating or preventinga nasal symptom of ocular allergy, comprising administering to the noseof a patient an effective amount of a compound of Formula II, itspharmaceutically acceptable salts, its N-oxides, hydrates, solvates,polymorphs, pro-drugs, or mixtures thereof. The terms a compound ofFormula II, patient, pharmaceutically acceptable, pharmaceuticallyacceptable salts, nasal symptoms, and effective amount all have theiraforementioned meanings and preferred ranges.

Further the invention includes a method of treating or preventing aclinical symptom of ocular allergy, comprising administering to the eyeof a patient an ophthalmic composition comprising a compound of FormulaII, its pharmaceutically acceptable salts, its N-oxides, hydrates,solvates, polymorphs, pro-drugs, or mixtures thereof. The terms acompound of Formula II, clinical symptom, patient, ophthalmiccomposition, pharmaceutically acceptable, and pharmaceuticallyacceptable salts, all have their aforementioned meanings and preferredranges.

Further, still the invention includes a method of treating or preventinga mechanistic symptom of ocular allergy, comprising administering to theeye of a patient an ophthalmic composition comprising a compound ofFormula II, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof. Theterms a compound of Formula II, mechanistic symptom, patient,pharmaceutically acceptable, and pharmaceutically acceptable salts, allhave their aforementioned meanings and preferred ranges.

Still further, the invention includes a method of treating or preventinga clinical symptom of ocular inflammation, comprising administering tothe eye of a patient an ophthalmic composition comprising a compound ofFormula II, its pharmaceutically acceptable salts, its N-oxides,hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof. Theterms a compound of Formula II, clinical symptom, patient,pharmaceutically acceptable, and pharmaceutically acceptable salts,ocular inflammation, and ophthalmic composition all have theiraforementioned meanings and preferred ranges.

Yet still further, the invention includes a method of treating orpreventing a mechanistic symptom of ocular inflammation, comprisingadministering to the eye of a patient an ophthalmic compositioncomprising a compound of Formula II, its pharmaceutically acceptablesalts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, ormixtures thereof. The terms a compound of Formula II, mechanisticsymptom, patient, pharmaceutically acceptable, and pharmaceuticallyacceptable salts, ocular inflammation, ophthalmic composition all havetheir aforementioned meanings and preferred ranges.

Further, the invention includes an ophthalmic composition comprising acompound of Formula II, its pharmaceutically acceptable salts, itsN-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixturesthereof. The terms ophthalmic composition, a compound of Formula II,pharmaceutically acceptable salts all have their aforementioned meaningsand preferred ranges. It is preferred that said ophthalmic compositionfurther comprise a vehicle as defined herein.

Yet further still, the invention includes use of a compound of FormulaII, its pharmaceutically acceptable salts, its N-oxides, hydrates,solvates, polymorphs, pro-drugs, or mixtures thereof in the preparationof a medicament for the treatment or prevention of a clinical symptom ofocular allergy. The terms a compound of Formula II, clinical symptom,ocular allergy pharmaceutically acceptable salts all have theiraforementioned meanings and preferred ranges Yet still further, theinvention includes use of a compound of Formula II, its pharmaceuticallyacceptable salts, its N-oxides, hydrates, solvates, polymorphs,pro-drugs, or mixtures thereof in the preparation of a medicament forthe treatment or prevention of a clinical symptom of ocularinflammation. The terms a compound of Formula II, clinical symptom,ocular inflammation pharmaceutically acceptable salts all have theiraforementioned meanings and preferred ranges.

Yet further still, the invention includes use of a compound of FormulaII, its pharmaceutically acceptable salts, its N-oxides, hydrates,solvates, polymorphs, pro-drugs, or mixtures thereof in the preparationof a medicament for the treatment or prevention of a mechanistic symptomof ocular allergy or ocular inflammation. The terms a compound ofFormula II, mechanistic symptom, ocular allergy, ocular inflammation,pharmaceutically acceptable salts all have their aforementioned meaningsand preferred ranges.

Still further, the invention includes a kit comprising an ophthalmiccomposition comprising a compound of Formula II, its pharmaceuticallyacceptable salts, its N-oxides, hydrates, solvates, polymorphs,pro-drugs, or mixtures thereof contained within a container preparedfrom a pharmaceutically acceptable packaging material. The termsophthalmic composition, alcaftadine, pharmaceutically acceptable saltsand pharmaceutically acceptable packaging material, all have theiraforementioned meanings and preferred ranges.

EXAMPLES

The invention is further demonstrated in the following examples. Theexamples are for purposes of illustration and are not intended to limitthe scope of the present invention.

Example 1 Alcaftadine Ophthalmic Solutions

Ophthalmic solutions containing alcaftadine were prepared in accordancewith Table I. To assure sterility, the solutions were passed through0.22 micron sterilizing filter prior to being filled into LDPE bottlesthat had previously undergone ethylene oxide sterilization. TABLE 1Conc. Conc. Conc. INGREDIENT mg/mL mg/mL mg/mL Alcaftadine 1.0 2.5 5.0Dibasic sodium phosphate USP 11.1  11.1  11.1  Monobasic potassiumphosphate NF 5.1 5.3 5.6 Sodium chloride USP 2.4 2.3 2.1 Benzalkoniumchloride NF 0.2 0.2 0.2 Edetate disodium USP 1.1 1.1 1.1 Purified waterUSP Q.S. Q.S. Q.S.

Example 2 Anti-Allergic Activity

The effect of alcaftadine against acute-phase reactions of allergicconjunctivitis (edema and erythema) was compared to the effect of otherknown anti-allergens in guinea pigs that were systemically sensitized torabbit skin squames and topically challenged 17 days later with rabbitallergens.

Anesthetized male albino guinea pigs (Dunkin-Hartley) weighing about 230to 250 g were injected intramuscularly at the left quadriceps with 50 μlof purified rabbit allergens. The rabbit allergens consisted ofAl(OH)₃-adsorbed rabbit squames (Halab, Brussels, Belgium), which hadbeen homogenized and washed clean of preservative (0.5% (V/V) phenol)with sterile physiologic saline.

Alcaftadine was orally administered to each eye in dosages ranging from0.005 mg/kg to 1.0 mg/kg at 24 hours and 1 hour before challenge. Othertest compounds included oxatomide, ketotifen, astemizole, cetirizine,loratadine and terfenadine, administered at dosages of 0.1 mg/kg and 1.0mg/kg.

At day 17 post sensitization the left eye was challenged by instilling25 μl of 100% normal rabbit serum. At the same time as the allergenchallenge, the right eye was instilled with 25 μl of 1.5 mg/ml histaminedihydrochloride (98%, Sigma) dissolved in deionised, Millipore-filteredwater.

Thirty minutes after challenge, edema and erythema were assessed in thetarsal and bulbar conjunctiva of both eyes and scored as absent (0),weak (1), moderate (2), severe (3), or very severe (4) by a trainedtechnician. Alcaftadine significantly alleviated the acute allergicsymptoms beginning at doses of 0.1 mg/kg. In this test, alcaftadine wasfound to be more potent (on an equivalent mg/kg basis) than oxatomide,ketotifen and terfenadine and significantly more potent than astemizole,cetirizine and loratadine.

Example 3 Activity of Topical Alcaftadine in Allergic Conjunctivitis

Alcaftadine has been shown to prevent signs and symptoms of allergicconjunctivitis in a murine model of active anaphylaxis.

Male SWR/J mice (The Jackson Laboratory, Bar Harbor, Me.), aged 5-7weeks and weighing between 12.55 and 17.73 grams, were sensitized with a100 μl dose of a suspension of 50 μg short ragweed allergen (Greer Labs,Inc., Lenoir, N.C.) and 1 mg of aluminum hydroxide (Fisher Scientific,Pittsburgh, Pa.) by intraperitoneal injection at two weeks and prior totreatment and challenge.

On Day 14, the mice were first given baseline exams to ensure that theydid not present with significant irritation prior to treatmentadministration and challenge. The mice were then dosed topically in theeye with an alcaftadine 0.0625% ophthalmic solution, positive control(ophthalmic solution comprising combination of ketotifen 0.05% andpheniramine 0.5%), or placebo prior to an ocular challenge with 1.5 mgshort ragweed allergen in phosphate buffered saline.

Fifteen (15) minutes after allergen challenge, mice were evaluated forclinical signs of allergic conjunctivitis by scoring for conjunctivalredness, chemosis, tearing, and lid edema. Severity of clinical signswas scored by a trained technician using a standardized 0-2 scale.Alcaftadine was more effective than the positive control in preventingitching, redness, chemosis and lid edema. alcaftadine was as effectiveas the positive control and more effective than placebo in preventingtearing.

Example 4 Effects of Alcaftadine Ophthalmic Solution in Humans-

The anti-allergic effect of a single dose of each of the threeconcentrations of alcaftadine ophthalmic solutions from Example 1 wasassessed in a Conjunctival Allergen Challenge (“CAC”) performed on adultvolunteers with a history of allergic conjunctivitis. Subjects wereselected after two visits to confirm reactivity to the allergenchallenge. Subjects qualified for the study if they had a positive skintest and ocular reaction to at least one of several common allergenssuch as cat hair, cat dander, tree pollen, grass pollen, and the like.Subjects were then challenged with allergen by instilling reconstitutedcommercially available allergen into each eye on two separate visits 16hours and 15 minutes after the bilateral instillation of alcaftadine,PATANOL® olopatadine hydrochloride ophthalmic solution 0.1% (Alcon,Inc., Forth Worth, Tex.) or vehicle, and the clinical response assessed.

Prevention of Ocular Itching

Patients administered alcaftadine 16 hours prior to challenge exhibiteda dose-related inhibition of ocular itching. All concentrations ofalcaftadine showed lower mean itching scores based on subject evaluationusing a 5 point scale (i.e., less itching) than vehicle or PATANOL at 3,5 and 7 minutes post-challenge. When challenged 15 minutespost-treatment, the alcaftadine-treated subjects exhibited adose-related inhibition of ocular itching as compared to placebo, andthe 0.25% treatment group had lower itching scores than vehicle orPATANOL.

Prevention of Conjunctival Redness

Patients administered alcaftadine 16 hours prior to challenge alsoexhibited a dose-related inhibition of conjunctival redness. Assessmentswere made at 7, 15 and 20 minutes post challenge based on investigatorevaluations of redness using a 5 point scale. All concentrations ofalcaftadine showed various degrees of reduction in mean conjunctivalredness scores at most assessment times. When challenged at 15 minutespost-treatment, the 0.25% treatment group had lower scores than vehicleor PATANOL.

Prevention of Nasal Symptoms

Similar results were observed for the prevention of nasal symptomsinduced by CAC. Nasal symptoms of sneezing, rhinorrhea, pruritis (nasaland ear/palate) and nasal congestion were assessed by subjects usingstandardized scales.

All concentrations of alcaftadine showed some degree of relief of nasalcongestion and rhinorrhea at various time points. These numbers oftenreached statistical significance versus either placebo and in someinstances active control (p≦0.05). Similar results were demonstrated forthe parameters of itching in the nose, palate and ear. No effect onsneezing was demonstrated in this study; however, baseline sneezingincidence (pre-treatment) was likely too low to detect a therapeuticeffect if one existed. In summary, significant reduction of rhinorrhea,nasal congestion, and pruritis of the palate/ear were seen at majorityof timepoints at both onset and duration visits.

Example 5 Effect of Sterilization with Gamma Irradiation

Alcaftadine is susceptible to oxidation and the primary degradationproduct has been identified as the N-oxide structure below. Theexistence of the N-oxide structures as the primary oxidative degradationpeak was confirmed as having the same HPLC relative retention time as asynthetically produced N-oxide structures and also confirmed using massspectroscopy.

To investigate the effects of the gamma irradiation process used tosterilize the LDPE bottles, the chemical stability of alcaftadine wasstudied using different lots of bottles, either gamma irradiated ornon-sterilized.Materials and Methods:

-   -   The following materials were used in this study. 2.5 mg/ml        alfactadine solution    -   Non-sterilized 5 ml LDPE bottles (DUPONT 20-6064) from Bünder        Glas GmbH (Germany)    -   Non-sterilized 5 ml LDPE bottles (DUPONT 20-6064) from Rexam        (France)    -   Sterilized (gamma irradiated at 25 kGy) LDPE bottles (DUPONT        20-6064) from Bünder Glas GmbH    -   Scintillation vials—20 ml clear glass with Teflon coated cap        Millex GV syringe filter unit        Results:

Chemical stability results for alcaftadine solution stored in eithernon-sterilized bottles (Rexam or Bunder Glas) or gamma-irradiatedbottles (Bunder Glas) are summarized in Table 2. All samples wereassayed by HPLC after 6 and 14 days of storage at 50° C. Light exposurewas not controlled during storage. The solution was stored in eitherRexam bottles (without sterilization), Bunder Glas bottles (sterilizedwith y Irradiation or without sterilization) and in glass vials. TABLE 2N- Container Sterilization Storage Time Alcaftadine Oxide DescriptionProcess Condition (days) (mg/mL) (%) Initial N/A N/A NA 2.47 0.02Solution Glass Vials N/A  5° C. 6 2.52 0.008 Glass Vials N/A 50° C. 62.57* 0.024 Rexam None 50° C. 6 2.46 0.015 14 2.48 0.033 Bunder GlasNone 50° C. 6 2.48 0.018 14 2.48 0.042 Bunder Glas Y 50° C. 6 2.41 0.199Irradiation 14 2.39 0.202*Sample showed significant evaporation, consistent with the higher assayvalue observedConclusions

Alcaftadine stored in gamma irradiated Bunder Glas bottles showedsignificantly increased levels of N-oxide formation compared with glassvials and non-sterilized Rexam and Bunder Glas bottles. This datasuggests that the gamma irradiation sterilization process may be theprimary cause of oxidation by inducing chemical or physical changes inthe polyethylene bottle.

Example 6 Effect of Sterilization with Ethylene Oxide

Both non-sterilized Rexam and Bunder Glas bottles were sterilized usingethylene oxide. Stability studies were initiated and ethylene oxidelevels determined to be <1 ppm (under ambient conditions) using a waterextraction test method (ANSI/AAMI/ISO: 10993-7) by AppTec (Marietta,Ga.).

Materials and Methods:

The 2.5 mg/ml alcaftadine solution was used in this study. Bottles weresterilized using ethylene oxide in accordance with the protocol setforth in Table 3: TABLE 3 Phase Parameter Set-point PRECONDITIONINGTemperature 40° C. % Rel. Humidity 60% Time ≧16 hours Transfer time → 45minutes sterilizer ETO processing Temperature 45° C. Initial vacuumPressure 70 mbarA Steam injection Pressure 42 mbars Steam dwell Time 45minutes ETO exposure Nitrogen injection 250 mbarA ETO injection 744mbarA Gas dwell time 3 Hours ETO concentration 800 ± 50 mg/l Gas removal(calculated) 75 mbarA 2 nitrogen washes Pressure 75 → 500 → 75 mbarA 1air wash Pressure 75 → 500 → 75 mbarA Pressure Aeration Temperature 40°C. Time 10 daysResults:

The chemical stability results for alcaftadine stored in ethylene oxidesterilized Rexam and Bunder Glas bottles are summarized in Table 4.TABLE 4 Time (days) Container Sterilization at N-Oxide DescriptionProcess 50° C. Alcaftadine (mg/ml) (%) Glass Vials N/A 5 2.48 0.022 142.51 0.028 Bunder Glas Ethylene Oxide 5 2.46 0.052 14 2.48 0.074 RexamEthylene Oxide 5 2.50 0.018 14 2.58 0.025

Conclusions: Based on 14 days of storage at 50° C. under ambient lightconditions, alfactadine has significantly lower N-oxide levels inethylene oxide sterilized Bunder Glas bottles than gamma irradiatedbottles. In this study, the amount of N-oxide formation in Rexam bottlesis similar to that observed in the glass vials (0.025% and 0.028%,respectively) and slightly higher in the Bunder Glas bottles (0.074%)after 14 days of storage at 50° C.

Example 7 Effect of Alcaftadine on Mast Cell Degranulation

The cell stabilization potential of alcaftadine was evaluated using theRBL-CCR1 (Rat Basophil Leukemia-Chemokine Receptor-1) cell line as a wayof investigating its ability to effect mast cell stabilization. The useof basophil cell lines to assess anti-allergic drugs for cellstabilization potential is well established. Physiologically, basophilsare similar to mast cells, containing pre-formed inflammatory mediatorsthat are released through a similar degranulation process involving IgEcross-linking. Because these cell lines are readily available, theypresent efficiencies over conducting stabilization assays in mast celllines.

Cultured RBL-CCR1 cells were sensitized to anti-DNP IgE. Followingsensitization, cells were treated with various concentrations ofalcaftadine or its major active metabolite (0.083%, 0.0083% and0.00083%) or ophthalmic vehicle (placebo), and stimulated with DNPand/or MIP-1a( Macrophage Inflammatory Protein-1 alpha, a degranulationenhancer) to induce degranulation. A 10× and 100× placebo controlsolution (ophthalmic vehicle solution) in D-DEM (Dulbecco's modifiedEagle's medium) was also employed.

Stimulation with DNP-HSA (2,4-Dinitrophenyl hapten conjugated to HumanSerum Albumin) alone and MIP-1α alone did not induce significant levelsof degranulation. However, co-stimulation induced a robust degranulationresponse. With all three degranulation stimuli, (DNP-HAS alone,MIP-1αalone, and co-stimulation) the highest concentration of parentdrug, metabolite, and placebo caused unanticipated and significantdegranulation,

In the co-stimulation assay, alcaftadine concentrations of 0.0083% and0.00083% were superior at cell stabilization compared to co-stimulationcontrol and negative controls (placebo 10× [vehicle 3.3% in D-MEM] andplacebo 100× [vehicle 0.33% in D-MEM] respectively). This superioritywas statistically significant (p<0.0001 for both concentrations).Treatment with6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) at concentrations of 0.0083% and 0.00083% wasalso superior at cell stabilization compared to co-stimulation controland negative controls (placebo 10× and placebo 100×, respectively).Again, statistical significance was achieved (p<0.0001 for bothconcentrations). Treatment with placebo 10× alone also indicated somestabilization compared to the co-stimulation control and the effect wasstatistically significant (p<0.05).

The addition of the alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) enhanced the stabilization effect at the 10×dose level, demonstrating the positive effect of the test agents atmembrane stabilization. Stabilization was not observed at the 100×dilution level. Overall, the results of this study suggest thealcaftadine and6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) are effective membrane stabilizing agents.

Example 8 Effect of Alcaftadine on Human H₄ Receptor

The pharmacological activity of alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) on the human histamine H₄ receptor (H₄R) wasinvestigated. The H₄R is the fourth histamine receptor that has beenidentified and it appears to be primarily expressed on eosinophils, Tcells, dendritic cells, basophils and mast cells, cell types intimatelyinvolved with development and perpetuation of allergic responses. H₄Rhas been shown to mediate mast cell, eosinophil and dendritic cellchemotaxis and can effect cytokine production from dendritic cells and Tcells. Antagonists for the receptor are clearly anti-inflammatory invivo and are efficacious in animal models of asthma and colitis.Alcaftadine and6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) were tested using cells transfected with thereceptor for binding to the H₄R and for an indication as to whether theywere agonists or antagonists of the receptor.

Alcaftadine and6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) were prepared at 10 mM in 100% dimethylsulfoxide(DMSO) for the binding assays and at 10 mM in Na/K phosphate buffer, pH7.0, for the cellular assays. Cell pellets from SK-N-MC cellstransfected with human H₄ receptor were homogenized in 20 mMTris-HCl/0.5 mM ethylendediaminetetraacetic acid (EDTA) pH 8.0 (TEbuffer). Supernatants collected after centrifugation at 800 g wererecentrifuged at 30,000 g for 30 min. Pellets were re-homogenized in TEbuffer. For competition binding studies, membranes were incubated with10 nM [³H]histamine with or without test compounds for 45 min at 25° C.Non-specific binding was defined with 100 μM cold histamine. Ki valueswere calculated based on an experimentally determined K_(d) value of 5nM for [³H]histamine and a ligand concentration of 10 nM according toCheng and Prusoff. Seven concentrations of compound were tested spanning10⁻¹¹ to 10⁻⁵ M with each concentration being run in triplicate. Thetriplicates were averaged and an IC₅₀ (50% inhibitory concentration)curve was generated. This assay was run twice and the results arereported as the average of the two runs.SK-N-MC cell lines were createdthat express a reporter gene construct and the human H₄ receptorfull-coding region. The reporter gene was β-galactosidase under thecontrol of cyclic adenosine monophosphate (cAMP) responsive elements.Cells were plated in 96-well plates the night before the assay.Antagonists were added 10 min prior to the addition of histamine, whichwas added directly to the cell medium. Forskolin (5 μM finalconcentration) was added 10 min after the addition of histamine. Cellswere returned to the incubator for 6 h at 37° C. The medium was thenaspirated and cells were lysed with 25 μL of 0.1× assay buffer (10 mMsodium phosphate, pH 8, 0.2 mM MgSO₄, 0.01 mM MnCl₂) and incubated atroom temperature for 10 min. Cells were then incubated for 10 min with100 μL of 1× assay buffer containing 0.5% Triton and 40 mMβ-mercaptoethanol. Color was developed using 25 μL of 1 mg/mL substratesolution (chlorophenol red β-D-galactopyranoside; Roche MolecularBiochemicals, Indianapolis, Ind.). Color was quantitated on a microplatereader at absorbance 570 nm. For agonist determination a titration ofcompounds from 10⁻¹¹ to 10⁻⁴ M in duplicate were added in the absence ofhistamine. The values for the duplicates were averaged and used tocalculate the EC₅₀ (effective concentration 50) for the inhibition ofcyclic AMP production by forskolin. This assay was repeated three times.For antagonist determination a titration of histamine from 10⁻¹⁰ to 10⁻³M was run in duplicate in the presence of 1.2, 3.7, 11, 33, and 100 μMcompound. The duplicates were averaged and the EC₅₀ for histamine ateach of the different concentrations of compound were used for a Schildplot to derived the pA₂ values, which are the negative log of theconcentration of compound needed to shift the histamine EC₅₀ by 2-fold.

The alcaftadine and6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) were tested for their ability to displace[³H]-histamine binding to membranes from SK-N-MC cells stablelytransfected with histamine H₄ receptor. Competition with [³H]-histamineindicates that the compounds can bind to the receptor. The bindingcurves showed that alcaftadine binds to the receptor with an averageK_(i) value of 2.9 μM, whereas6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) does not bind to the receptor at concentrationsup to 10 μM.

To test whether alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) were H₄ receptor agonists, functional assayswere carried out in SK-N-MC cells transfected with the human histamineH₄ receptor. The ability of the compounds to inhibit forskolin-inducedcAMP increases was assessed. The results showed that histamine is anagonist of the receptor and causes a dose-dependent inhibition offorskolin-induced cAMP levels. However, neither alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) showed any inhibition of cAMP levels andtherefore neither is an agonist of the H₄ receptor at concentrations upto 100 μM.

To test whether alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) were antagonists of the H₄ receptor, the abilityof the compounds shift the EC₅₀ of the histamine inhibition offorskolin-induced cAMP increases was assessed in SK-N-MC cellstransfected with the human histamine H₄ receptor. The results showedthat increasing concentrations of alcaftadine caused parallel andrightward shifts in the histamine (HA) dose response curves leading toan increase in the EC₅₀ for histamine modulation of the H₄ receptor.This effect indicates that alcaftadine is a competitive antagonist ofthe receptor. The x-intercept of the Schild plot gives a pA2 value of5.6, which represents the negative log of the concentration ofantagonist need to induce a 2-fold shift in the histamine EC₅₀.Theoretically the pA2 value should be equal to the pK_(i), which isindeed observed (5.6 versus 5.5). Alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) did not cause any shift at concentrations up to100 μM, which is consistent with its inability to bind to the receptor.

These results indicate that alcaftadine binds to the H₄ receptor with anaverage K_(i) value of 2.9 μM. Neither alcaftadine or6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) is an agonist of the H₄ receptor atconcentrations up to 100 μM. However, alcaftadine, but not alcaftadineor6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid (CAS #147083-93-0) (up to 100 μM), is an antagonist of the humanhistamine H₄ receptor with a K_(i) value of 2.9 μM and pA₂ value of 5.6.

Example 9 Effects of Alcaftadine on the Integrity of ConjunctivalEpithelium

To investigate the ability of alcaftadine to maintain the integrity ofconjunctival epithelium, changes in the expression of the tight junctionproteins ZO-1 and E-cadherin (an induction for E-cadherin andqualitative change-from focal to diffuse for ZO-1) were evaluatedfollowing specific conjunctival allergen challenge. These changes areassociated with an increased permeability of the conjunctival and otherepithelial tissues. This in vivo experiment examined what effectalcaftadine (topical administration 1 and 2 hours pre-challenge) had onthe modulation of these proteins.

Mice were sensitized with short ragweed (SRW) in aluminum hydroxide viaintraperitoneal administration (Day 0, 7, and 15) and eye dropinstillation (Day 8 and 15). On Day 20, mice were further sensitizedwith eye drops containing only SRW. On Day 27, the mice were treatedtopically with either 5 μL alcaftadine or vehicle at 1 and 2 hours priorto challenge. SRW was instilled topically to both eyes. The expressionof these proteins in naive eyes were also monitored. Changes in theexpression of the tight junction proteins ZO-1 and E-cadherin wereobserved 1-hour post conjunctival allergen challenge with SRW. Theproteins were detected using FITC-conjugated monoclonal antibodiesspecific for ZO-1 and E-cadherin and were visualized by confocalmicroscopy (Ziess). Naïve eyes were from mice that had not beensensitized or challenged.

The naive (no challenge and no treatment) ZO-1 and E-cadherin proteinsshowed focal qualitative properties. However, comparing this to vehicletreated proteins, the vehicle treated proteins showed diffusequalitative changes. The transition from focal to diffuse staining isassociated with increased permeability of the epithelium. Comparingnaïve ZO-1 and E-cadherin proteins with alcaftadine-treated proteins,the results show minimal difference in qualitative properties. Images ofZO-1 and E-cadherin proteins showed no or minimal difference betweennaïve (negative control) and alcaftadine treatment, while there was aclear and distinct difference with vehicle treatment.

These results suggest alcaftadine maintains the integrity ofconjunctival epithelial tight junctions (typified by focal ZO-1expression) and inhibits the induction of E-cadherin expression normallyobserved following specific conjunctival allergen challenge.

Example 10 Effect of Alcaftadine on Vascular Leakage and CellularInfiltrates

Allergic inflammation can be separated into two distinct phases, theearly phase and the late phase. The early phase inflammatory responseoccurs rapidly following mast cell degranulation and is characterized byvascular endothelial cell gaping and leakage (i.e. swelling) anditching. The late phase inflammatory response peaks approximately 24hours after mast cell degranulation and is characterized by theappearance of cellular infiltrates (eosinophils and neutrophils). Botheosinophils and neutrophils are known to have a profound effect onexponentiating the late phase of the inflammatory response. Uponarriving at the site of inflammation, these cells release variousperoxidases and other antimicrobial factors that function to kill offinvading pathogens but, in the case of severe or chronic inflammation,damage surrounding tissues and cause further release of pro-inflammatorymediators.

Alcaftadine ophthalmic solution was evaluated to determine if topicaltreatment could reduce early phase and late phase allergic inflammationin a murine model of allergic conjunctivitis. Evans Blue Dye Leakage wasused to assess the impact on vascular permeability during the earlyphase inflammatory response Conjunctiva was taken at 24 hours, stained,and assessed for presence of neutrophil and eosinophil infiltration toevaluate late phase anti-inflammatory activity.

All mice were sensitized with short ragweed (SRW) in aluminum hydroxidevia intraperitoneal administration (Day 0, 7, and 14) and eye dropinstillation (Day 8 and 15). On Day 20, mice were again sensitized witheye drops of SRW.

On Day 27, two topical administrations of the clinical dose ofalcaftadine 2.5 mg/ml or vehicle were instilled at 1 and 2 hourspre-challenge. Alcaftadine and vehicle were administered topically at a5 μL saturating dose as treatment arms based on a randomization code.The treatment arms were as follows:

-   -   1. Negative Control. Sensitized, No challenge, no treatment        (negative control) (N=12)    -   2. Sensitized, Challenge, no treatment (positive control) (N=12)    -   3. Sensitized, Challenge, alcaftadine 2.5 mg/ml treatment (N=10)    -   4. Sensitized, Challenge, vehicle treatment (N=12)        Vascular leak was determined by means of Evans Blue Dye        Extravasation following allergen challenge for N=6 animals        except the alcaftadine treatment arm, where N=4 animals were        sacrificed and tested (two animals died during the procedure).        The treatment groups were compared using T-test and p<0.05 was        considered to be statistically significant. Dissection was        performed on the remaining animals, N=6 per treatment arm, 24        hours post-conjunctival allergen challenge (CAC) to assess        eosinophil and neutrophil recruitment in the forniceal area. The        tissues were processed either as frozen or plastic blocks prior        to sectioning on a microtome. The numbers of eosinophils and        neutrophils were determined both by light microscopy of Giemsa        or H&E stained sections or by immunohistochemistry using cell        specific monoclonal antibodies. The treatment groups were        compared using T-test and p<0.05 was considered to be        statistically significant.

There was no significant difference between the Negative Control (nochallenge group—Group 1) and Challenged but untreated (Group 2) group.However, as expected, there was a numerical difference with Group 2having higher scores of vascular leak. There was a significantdifference between the vehicle treatment (Group 4) and alcaftadinetreatment (Group 3) (p<0.05) groups with alcaftadine successfullypreventing vascular leakage compared to vehicle. As expected, there wasa significant difference between the negative control (no challengegroup-Group 1) and untreated but challenged group (Group 2). Alcaftadine(Group 3) did not prevent eosinophil or neutrophil recruitment comparedto controls.

Regarding vascular leakage, the typical induction of vascular leak wasnot observed in Group 2 (Challenged and untreated). However, alcaftadinedid significantly inhibit vascular leak following CAC as compared tomice treated with vehicle alone. Based on observations from previousstudies, the low level of vascular leak in Group 2 is atypical and ismost likely due to experimental error associated with in vivoexperiments. The decreased vascular leak in the alcaftadine treatedanimals is supportive of a therapeutic role for alcaftadine in thisconjunctival allergen challenge model.

In the evaluation of eosinophil or neutrophil recruitment, thesignificant difference between the negative and positive controlssuggests the model worked as expected. However, at the dose used,alcaftadine does not inhibit eosinophil or neutrophil recruitment in thelate phase response, in the murine model of allergic conjunctivitisfollowing allergen challenge in sensitized mice.

The results of this study indicate that alcaftadine did significantlyinhibit vascular leak following conjunctival allergen challenge.However, it does not appear to inhibit eosinophil or neutrophilrecruitment in the late phase response, in the murine model of allergicconjunctivitis following allergen challenge in sensitized mice.

Example 11 Systemic Levels of Alcaftadine in Patients

Patients were bilaterally dosed with 0.25% ophthalmic solutions ofalcaftadine for seven days. The plasma levels of those patients wasassessed pre-dose, and 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 18 hourpost medication instillation on days one and seven. Plasmaconcentrations of Alcaftadine reached C_(max) rapidly and declined tobelow the lower limit of quantification (0.01 ng/mL ) by 3 hours postdosing. Mean C_(max) values (highest value measured at any time point)were quite low, mean of 0.051 ng/mL on Day 1 and 0.060 ng/mL on Day 7;the maximum plasma concentrations were less than 0.12 ng/mL for allsubjects.

Example 12 Systemic Levels of6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid in Patients

Patients were bilaterally dosed with 0.25% ophthalmic solutions of6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid for seven days. The plasma levels of those patients was assessedpre-dose, and 0.25, 0.5, 1, 1.5, 2, 3, 4, 6, 8, 12, and 18 hour postmedication instillation on days one and seven. Plasma concentrations of6,11-dihydro-11-(1-methyl-4-piperdinylidene)-5H-imidazo[2,1-b][3]benazepine-3-carboxylicacid reacheached C_(max) rapidly and declined to below the lower limitof quantification (0.1 ng/mL) by 12 hours post dosing. Mean C_(max)values of 3.228 ng/mL on Day 1 and 2.715 ng/mL on Day 7; the maximumplasma concentrations was 7.23 ng/mL.

The entire disclosure of each patent, patent application, andpublication cited or described in this document is hereby incorporatedby reference.

Those skilled in the art will appreciate that numerous changes andmodifications can be made to the preferred embodiments of the inventionand that such changes and modifications can be made without departingfrom the spirit of the invention. It is, therefore, intended that theappended claims cover all such equivalent variations as fall within thetrue spirit and scope of the invention.

1. A method of treating or preventing a clinical symptom of ocular allergy, comprising administering to the eye of a patient an effective amount of alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 2. The method of claim 1 wherein the ocular allergy is allergic conjunctivitis.
 3. The method of claim 1 wherein said clinical symptom is selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, ear/palate prurtis, and sneezing.
 4. The method of claim 1 wherein said clinical symptom is ocular itching
 5. The method of claim 1 wherein said clinical symptom is ocular redness.
 6. The method of claim 1 further comprising treating or preventing at least two clinical symptoms.
 7. The method of claim 6 wherein said at least two clinical symptoms are ocular itching and ocular redness
 8. The method of claim 1, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 9. The method of claim 1 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 10. The method of claim 1 wherein the effective amount is between about 0.075 mg and about 0.125 mg.
 11. The method of claim 1 comprising administering an effective amount of alcaftadine.
 12. The method of claim 11 wherein effective amount is between about 0.075 mg and about 0.125 mg.
 13. The method of claim 11 wherein the effective amount is between less than about 0.25 mg to greater than about 0.015 mg.
 14. The method of claim 11 wherein the clinical symptom is ocular itching.
 15. The method of claim 11 wherein the clinical symptom is ocular redness
 16. The method of claim 11 further comprising treating or preventing at least two clinical symptoms.
 17. The method of claim 16 wherein said at least two clinical symptoms are ocular itching and ocular redness
 18. The method of claim 11, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 19. A method of treating or preventing a clinical symptom of ocular inflammation, comprising administering to the eye of a patient an effective amount of alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 20. The method of claim 19 wherein said clinical symptom is selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, pruritis (nasal and ear/palate), and sneezing.
 21. The method of claim 19 wherein said clinical symptom is ocular itching
 22. The method of claim 19 wherein said clinical symptom is ocular redness.
 23. The method of claim 19 further comprising treating or preventing at least two clinical symptoms.
 24. The method of claim 19, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 25. The method of claim 19 wherein effective amount is between less than about 7.1 μg/kg per day and about 3.5 μg/kg per day.
 26. The method of claim 19 wherein the effective amount is between less than about 0.25 mg to greater than about 0.015 mg.
 27. The method of claim 19 comprising administering an effective amount of alcaftadine.
 28. The method of claim 27 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 29. The method of claim 27 wherein the effective amount is between about 0.075 mg and about 0.125 mg.
 30. The method of claim 27 wherein the clinical symptom is ocular itching.
 31. The method of claim 27 wherein the clinical symptom is ocular redness
 32. The method of claim 27 further comprising treating or preventing at least two clinical symptoms.
 33. The method of claim 32 wherein said at least two clinical symptoms are ocular itching and ocular redness
 34. The method of claim 27, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 35. A method of treating or preventing a mechanistic symptom associated with ocular allergy or ocular inflammation comprising administering to the eye of a patient an effective amount of alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 36. The method of claim 35 wherein said mechanistic symptom is selected from the group consisting of vascular leakage, a reduction in the integrity of the conjunctival epithelial tight junctions, modulation of the H₄ receptor, and mast cell degradation.
 37. The method of claim 35 comprising administering an effective amount of alcaftadine.
 38. The method of claim 37 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg
 39. The method of claim 37 wherein the effective amount is between about 0.075 mg and about 0.125 mg.
 40. A method of treating or preventing a nasal symptom of ocular allergy, comprising administering to the nose of a patient an effective amount of alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 41. The method of claim 40 wherein the nasal symptom of allergy is selected from the group consisting of nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, and sneezing.
 42. The method of claim 41 further comprising treating or preventing at least two nasal symptoms of allergy.
 43. The method of claim 41 comprising administering an effective amount of alcaftadine.
 44. The method of claim 43 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 45. The method of claim 43 wherein the effective amount is between between about 0.075 mg and about 0.125 mg.
 46. A method of treating or preventing a clinical symptom of ocular allergy, comprising administering to the eye of a patient an ophthalmic composition comprising alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 47. The method of claim 46 wherein the ocular allergy is allergic conjunctivitis.
 48. The method of claim 46 comprising administering an ophthalmic composition comprising alcaftadine.
 49. The method of claim 48 wherein the ophthalmic composition comprises from about 0.005% to about 10% by weight of alcaftadine.
 50. The method of claim 48 wherein the ophthalmic composition comprises from about 0.2% to about 0.35% by weight of alcaftadine.
 51. The method of claim 48 further comprising treating or preventing at least two clinical symptoms of ocular allergy selected from the group consisting of of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, ear/palate pruritis, and sneezing.
 52. An ophthalmic composition comprising alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 53. The ophthalmic composition of claim 52, further comprising a vehicle.
 54. The ophthalmic composition of claim 52 comprising alcaftadine and a pharmaceutically acceptable vehicle.
 55. The ophthalmic composition of claim 52 wherein the ophthalmic composition comprises from about 0.005% to about 10% by weight of alcaftadine.
 56. The ophthalmic composition of claim 52 wherein the ophthalmic composition comprises from about 0.2% to about 0.35% by weight of alcaftadine.
 57. A kit comprising an ophthalmic composition comprising alcaftadine, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof contained within a container prepared from a pharmaceutically acceptable packaging material.
 58. The kit of claim 57 wherein the ophthalmic composition is in the form of an ophthalmic solution or suspension.
 59. The kit of claim 57 wherein the ophthalmic solution comprises between about 0.1% by weight and about 0.4% by weight of alcaftadine.
 60. The kit of claim 59 wherein the pharmaceutically acceptable packaging material is low density polyethylene or high density polyethylene.
 61. The kit of claim 59 wherein said container is sterilized with ethylene oxide prior of filing with said ophthalmic solution.
 62. A method of treating or preventing a clinical symptom of ocular allergy, comprising administering to the eye of a patient an effective amount of a compound of

its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 63. The method of claim 62 wherein the ocular allergy is allergic conjunctivitis.
 64. The method of claim 62 wherein said clinical symptom is selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, ear/palate prurtis, and sneezing.
 65. The method of claim 62 wherein said clinical symptom is ocular itching
 66. The method of claim 62 wherein said clinical symptom is ocular redness.
 67. The method of claim 62 further comprising treating or preventing at least two clinical symptoms.
 68. The method of claim 67 wherein said at least two clinical symptoms are ocular itching and ocular redness
 69. The method of claim 62, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 70. The method of claim 62 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 71. The method of claim 62 wherein the effective amount is between about 0.075 mg and about 0.125 mg.
 72. The method of claim 62 comprising administering an effective amount of a compound of Formula II.
 73. The method of claim 72 wherein effective amount is between about 0.075 mg and about 0.125 mg.
 74. The method of claim 72 wherein the effective amount is between less than about 0.25 mg to greater than about 0.015 mg.
 75. The method of claim 72 further comprising treating or preventing at least two clinical symptoms.
 76. The method of claim 75 wherein said at least two clinical symptoms are ocular itching and ocular redness
 77. The method of claim 72, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 78. A method of treating or preventing a clinical symptom of ocular inflammation, comprising administering to the eye of a patient an effective amount of a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 79. The method of claim 78 wherein said clinical symptom is selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, pruritis (nasal and ear/palate), and sneezing.
 80. The method of claim 79 further comprising treating or preventing at least two clinical symptoms.
 81. The method of claim 79, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 82. The method of claim 79 wherein the effective amount is between less than about 0.25 mg to greater than about 0.015 mg.
 83. The method of claim 79 comprising administering an effective amount of a compound of Formula II.
 84. The method of claim 83 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 85. The method of claim 83 wherein the effective amount is between about 0.075 mg and about 0.125 mg.
 86. The method of claim 83 further comprising treating or preventing at least two clinical symptoms.
 87. The method of claim 86 wherein said at least two clinical symptoms are ocular itching and ocular redness
 88. The method of claim 83, further comprising treating or preventing at least three clinical symptoms selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, and tearing
 89. A method of treating or preventing a mechanistic symptom associated with ocular allergy or ocular inflammation comprising administering to the eye of a patient an effective amount of a compound of Formula II, its pharmaceutically acceptable salts, N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 90. The method of claim 89 wherein said mechanistic symptom is selected from the group consisting of vascular leakage, a reduction in the integrity of the conjunctival epithelial tight junctions, and mast cell degradation.
 91. The method of claim 89 comprising administering an effective amount of a compound of Formula II.
 92. The method of claim 91 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg
 93. The method of claim 91 wherein the effective amount is between between about 0.075 mg and about 0.125 mg.
 94. A method of treating or preventing a nasal symptom of ocular allergy, comprising administering to the nose of a patient an effective amount of a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 95. The method of claim 94 wherein the nasal symptom of allergy is selected from the group consisting of nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, and sneezing.
 96. The method of claim 94 further comprising treating or preventing at least two nasal symptoms of allergy.
 97. The method of claim 94 comprising administering an effective amount of a compound of Formula II.
 98. The method of claim 97 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg.
 99. The method of claim 97 wherein the effective amount is between between about 0.075 mg and about 0.125 mg.
 100. A method of treating or preventing a clinical symptom of ocular allergy, comprising administering to the eye of a patient an ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 101. The method of claim 100 wherein the ocular allergy is allergic conjunctivitis.
 102. The method of claim 100 comprising administering an ophthalmic composition comprising a compound of Formula II.
 103. The method of claim 100 wherein the ophthalmic composition comprises from about 0.005% to about 10% by weight of a compound of Formula II.
 104. The method of claim 100 wherein the ophthalmic composition comprises from about 0.2% to about 0.35% by weight of a compound of Formula II.
 105. The method of claim 100 further comprising treating or preventing at least two clinical symptoms of ocular allergy selected from the group consisting of of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, ear/palate pruritis, and sneezing.
 106. A method of treating or preventing a mechanistic symptom of ocular allergy, comprising administering to the eye of a patient an ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 107. The method of claim 106 wherein said mechanistic symptom is selected from the group consisting of vascular leakage, a reduction in the integrity of the conjunctival epithelial tight junctions, and mast cell degradation.
 108. The method of claim 106 comprising administering an effective amount of a compound of Formula II.
 109. The method of claim 106 wherein effective amount is less than about 0.25 mg to greater than about 0.015 mg
 110. The method of claim 108 wherein the effective amount is between between about 0.075 mg and about 0.125 mg.
 111. A method of treating or preventing a clinical symptom of ocular inflammation, comprising administering to the eye of a patient an ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 112. The method of claim 111 wherein the ocular allergy is allergic conjunctivitis.
 113. The method of claim 110 comprising administering an ophthalmic composition comprising a compound of Formula II.
 114. The method of claim 110 wherein the ophthalmic composition comprises from about 0.005% to about 10% by weight of a compound of Formula II.
 115. The method of claim 110 wherein the ophthalmic composition comprises from about 0.2% to about 0.35% by weight of a compound of Formula II.
 116. The method of claim 110 further comprising treating or preventing at least two clinical symptoms of ocular allergy selected from the group consisting of ocular itching, ocular redness, swelling of the eyelids, chemosis, tearing, and nasal inflammation, nasal congestion, rhinorrhea, nasal pruritis, ear/palate pruritis, and sneezing.
 117. A method of treating or preventing a mechanistic symptom of ocular inflammation, comprising administering to the eye of a patient an ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 118. The method of claim 117 wherein said mechanistic symptom is selected from the group consisting of vascular leakage, a reduction in the integrity of the conjunctival epithelial tight junctions, and mast cell degradation.
 119. An ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof.
 120. The ophthalmic composition of claim 119, further comprising a vehicle.
 121. The ophthalmic composition of claim 119 comprising a compound of Formula II and a pharmaceutically acceptable vehicle.
 122. The ophthalmic composition of claim 121 wherein the ophthalmic composition comprises from about 0.005% to about 10% by weight of a compound of Formula II.
 123. The ophthalmic composition of claim 119 wherein the ophthalmic composition comprises from about 0.2% to about 0.35% by weight of a compound of Formula II.
 124. A kit comprising an ophthalmic composition comprising a compound of Formula II, its pharmaceutically acceptable salts, its N-oxides, hydrates, solvates, polymorphs, pro-drugs, or mixtures thereof contained within a container prepared from a pharmaceutically acceptable packaging material.
 125. The kit of claim 124 wherein the ophthalmic composition is in the form of an ophthalmic solution or suspension.
 126. The kit of claim 124 wherein the ophthalmic solution comprises between about 0.1% by weight and about 0.4% by weight of a compound of Formula II.
 127. The kit of claim 124 wherein the pharmaceutically acceptable packaging material is low density polyethylene or high density polyethylene.
 128. The kit of claim 127 wherein said container is sterilized with ethylene oxide prior of filing with said ophthalmic solution. 